Cell Biology and Toxicology

, Volume 7, Issue 1, pp 1–14

Co-cultures of hepatocytes with epithelial-like cell lines: Expression of drug-biotransformation activities by hepatocytes

  • M. Teresa Donato
  • José V. Castell
  • M. José Gómez-Lechón
Article

To improve long-term expression of drug biotransformation activities in hepatocytes, we have examined the suitability of several epithelial-like cell lines (MDCK, MS and L-132) for supporting functional co-cultures with rat hepatocytes. Cells were selected on the basis of their compatibility with hepatocytes, formation of stable monolayers in the absence of serum and lack of drug biotransformation activities. The expression of individual elements of the biotransformation system was evaluated in these co-cultures. Co-cultured hepatocytes remained viable and showed a characteristic polygonal shape for more than a week. Depending on the cell line used, levels of aryl hydrocarbon hydroxylase and 7-ethoxycoumarin O-deethylase activities of co-cultured hepatocytes oscillated between 24–47% of their initial value after 4 days in culture. The highest levels of monooxygenase activity were found in hepatocytes co-cultured with MS cells (41–47%). In contrast, these activities decreased to 6% when hepatocytes were maintained in pure culture for the same period. The activities of the conjugating enzymes UDP-glucuronyltransferase and glutathione S-transferase were maintained at nearly the initial levels during the complete period of study, both in pure and mixed-cultures, regardless of the cell line used. MS cells adapted themselves much better to serum-free culture conditions, and the co-culture with rat hepatocyte was technically easier. After one week, total cytochrome P450 and reduced glutathione in rat hepatocytes/MS co-cultures were 31% and 127% respectively of the day O values, whereas they were undetectable in pure culture. A clear induction of monooxygenase activities by methylcholanthrene, phenobarbital and ethanol could be observed by the 5th day in MS cells/hepatocyte co-cultures. The fact that the results of our work show the suitability of MS cells, an epithelial-derived cell line, for improving the expression of biotransformation enzymes of cultured hepatocytes opens new possibilities of simplifying co-cultures for their use in drug-metabolism studies.

Key words

co-cultures conjugating enzymes epithelial-like cell lines induction monooxygenases rat hepatocytes 

Abbreviations

AHH

aryl hydrocarbon hydroxylase

CDNB

1-chloro-2,4-dinitrobenzene

DMEM

Dulbecco's modified Eagle's medium

ECOD

7-ethoxycoumarin O-deethylase

EDTA

ethylenediamine tetraacetic acid

Et-OH

ethanol

GSH

reduced glutathione

GSH-t

glutathione S-transferase

MC

3-methylcholanthrene

PB

phenobarbital

UDP-Gt

UDP-glucuronyltransferase

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Copyright information

© Princeton Scientific Publishing Co., Inc 1991

Authors and Affiliations

  • M. Teresa Donato
    • 2
  • José V. Castell
    • 2
  • M. José Gómez-Lechón
    • 2
  1. 1.Unidad de Hepatología ExperimentalCentro de Investigación Hospital La Fe Servicio Valenciano de SaludValenciaSpain
  2. 2.Unidad de Hepatología ExperimentalCentro de Investigación, Hospital La Fe, Servicio Valenciano de SaludValenciaSpain

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